Manuscript copyright
Berezkina Elena Sergeevna
The effect of preconceptional alcohol intoxication in female rats on the containment of macro- and microelements in different organs of the progenies.
14.01.27 – Narcology
Autoabstract
Of the Ph.D. (Biology) thesis
Moscow – 2010
The reported investigation has been performed in the Institute of Bioelementology, a subunit of the State Educational Institution of Higher Professional Education «The Orenburg State University» (rector – D.Sc. (Economics), professor Kovalevskiy V.P.)
Research advisor:
Doctor of Medical Science, Skalnyi Anatoliy Viktorovich
professor
Official opponents:
Doctor of Biological Sciences Proskuryakova Tatyana Vasilievna
Doctor of Biological Sciences, Karganov Mihail Yurievich
professor
Leading organization: Peoples’ Friendship University of Russia
The Ph.D. defense is going to be held on «_____» ______2010at ______(time) during the Dissertation Council board (Ph.D. and M.D.) (session number 208.051.01, Federal State Institution “National Scientific Center”, a subunit of the Russian Healthcare Supervision Agency). The Council board address: 119002 Moscow, Malyi Mogiltsevskiy lane, facility 3.
The thesis materials are available in the Scientific Library of the Federal State Institution “National Scientific Center”, a subunit of the Russian Healthcare Supervision Agency, using the following address: 119002 Moscow, Malyi Mogiltsevskiy lane, facility 3.
The autoabstract has been sent on «_____» ______2010.
Academic secretary
Ph.D. (Biology) Lvova O.F.
The scientific investigation overall characteristics
The timeliness of the investigation. In recent years Russian population demonstrates a significantly higher level of women with abusive alcohol consumption (Koshkina Е.А. Kirzhanova V.V., 2008; Altshooler V.B., Mokhnachev С.О., 2008), including the abusive drinking behavior for the pregnancy period. Amongst all the consequences of the alcohol’s threat to the fetus the leading role belongs to the brain impairments detectable on the different stages of the postnatal period due to different cognitive disorders, including attention disorders, difficulties in acquiring information, mnemonic disorders and others. (Arzumanov Yu.L., 2001; Altshooler V.B., Kravchenko S.L., 2008).
A major alarming concern is that most people think that consuming low-alcohol beverages or consuming low amount of alcohol before getting pregnant is safe for the progeny. Such a fallacy is especially wide-spread among the teenagers and youngsters.
It is well-known that alcohol consumption leads to metabolic and functional disorders in almost all the systems of the organism (Ivanets N.N., 2008). The reported investigations sometimes show the influence of the alcohol intoxication on the isolated macro- and microelements within the organisms of the mother and children (Skalnyi А.V., 1990; Rosenbaum J.S., Zidenberg-Cherr S., Keen C.L. 1991; Keiver K. et al., 1997; Kohgo Y. et al., 2005; Cylwik B., Chrostek L., Szmitkowski M., 2008, etc.). However, these data are quite few and fragmental. The actual problem that is still unsolved is - are there any changes in mineral metabolism of a progeny in mothers that fully stopped consuming alcohol before deciding to have a baby?
The human organism contains a full spectrum of chemical elements (ChE.), and each one can be found in the surrounding environment. Some of them may be present in the organism for a short period of time (without taking part in the metabolic processes), but most of ChE are involved in different metabolic and physiologic functions. The activity spectrum of the macro- and microelements is quite wide and mostly involves multiple interactions with different enzymes, hormones, neuromediators, vitamins, peptides and other biologically active substances (Avtsyn А.P. et al, 1991; Tutelyan V.А. et al., 2002; Skalnaya M.G., Notova S.V., 2004; Oberlis D. et al., 2008). According to the opinion of several researchers, an impairment of the mineral metabolism could be one of the triggering mechanisms in alcohol intoxication (Skalnyi А.V., 1990; Novoselov Ya.B., 2001; Panchenko L.F. et al., 2004; Keen C.L. et al., 1997 etc.).
Several research reports show the effectiveness of pre-natal micronutrients status disorders prevention in infants, especially in case of pathologic processes within the mother’s organism (Zhuravleva E.A. et al., 2007; Molloy A.M. et al., 2008; Zeisel S.H., 2008 etc.). In particular – USA prevention program protocols for mothers that abuse alcohol, includes undergoing a correction course with folic acid and choline formulations intake and, which is important, with zinc-containing formulations intake (Keen C.L. et al., 2010). However, there are no available results of the investigational experiments concerning the antenatal metabolic correction treatment (macro- and microelements) in mature progenies of females that consumed alcohol before getting pregnant.
The current scientific data states that zinc formulations can be used to provide corrective treatment, as zinc deficiency notably takes place during alcohol intoxications, as well as in cases of embryo- and fetopathy due to alcohol abuse in mothers (during the pregnancy) (Semenov А.S., Skalnyi А.V., 2009; Maret W., Sandstead H.H., 2008). In addition to that, zinc deficiency is very often found in pregnant women and breast-feeding mothers (even without previous alcohol abuse), as well as it is often found in children (Oberleas D., 1996, Yasui M. et al., 1997, DiSilvestro R.A., 2005; Shah D., Sachdev H.P., 2006; Hess S.Y., King J.C., 2009 etc.), especially the ones with depressed immune system activity and various disorders of the central nervous system (Prasad A.S., 1983, 2009).
All the abovementioned information defines the actuality of the present experimental investigation.
Objective: To assess the effect of the alcohol intoxication in female rats before pregnancy by measuring the macro- and microelements concentrations within different structures of the brain and in different organs of the mature progenies.
Investigation goals:
1. To define the concentration of the macro- and microelements in the organs and brain tissue in mature progenies of the female rats after alcohol exposure before mating with intact (non-alcoholic) males.
2. To assess the effectiveness of zinc sulfate administration in rat females during different periods (mating, pregnancy, breast-feeding) by measuring the macro- and microelements concentration in brain tissue (different brain structures) samples obtained from the rat mature progenies.
3. To perform the significance assessment of the data concerning chemical elements proportions within the investigated brain structures and organs of the mature progenies, including the groups of female rats that were taking zinc sulfate.
Scientific novelty
This experiment is the first to obtain data concerning the concentrations of macro-elements (calcium (Ca), phosphorus (Р), potassium (K), sodium (Na), magnesium (Mg)) and microelements (iron (Fe), zinc (Zn), copper (Cu) and manganese (Mn)) in tissue samples obtained from organs and different brain structures of mature rat progenies (female rats were alcoholized before pregnancy) with further comparative analysis against a group of intact female rats).
For the first time it has been stated that alcohol administration to female rats (before mating with intact males) results in the disproportions in ChE concentrations within different organs and tissues, which leads to elevated levels of Ca, K and Na in the hepatic tissues, Р – in kidney tissues and К- in the cardiac muscle. The brain tissue samples (different segments of the brain) presented decreased levels of Na, Mg, К, Fe, Zn (brain cortex), Ca, P, Fe, Mn (white substance), Mn (hippocampus). The cerebellum decreased level of Mn is followed by an elevated concentration of Zn.
For the first time it has been reported that after an alcohol exposure in rat females (before mating) the most significant deviations of macro- and microelements concentrations (in mature progenies) take place in the hepatic tissue, the brain cortex and the brain white matter.
For the first time it has been shown that antenatal and postnatal administration of zinc sulfate (after an alcohol exposure) effectively corrects the alcohol impact on mother’s organism (alcohol exposure before pregnancy). A positive therapeutic effect of zinc sulfate administration is expressed in normalizing the Mn concentrations in the cerebellum and hippocampus tissues, K, Na, Mg, Fe, Zn – in the brain cortex samples, Ca, P, Fe, Mn – in the brain white matter samples (in mature rat progenies), thus, the observed effect can successfully prevent the development of pathological deviations, caused by the deficiency of the abovementioned chemical elements.
Propositions to be defended
1. Alcohol consumption cancellation (even after a short period of exposure) in rat females before mating with intact males, as well as for the pregnancy period and during breast-feeding, does not prevent the development of macro- and microelements disproportional deviations within the organs and brain structures (in mature progeny).
2. The most sensitive parameters (concerning the effect of the alcohol exposure in rat females) are the concentrations of chemical elements within the hepatic tissues, the brain cortex and white matter of the mature progenies.
3. Disproportional deviations of the chemical elements concentrations (Diselementosis) within the brain structures of the mature rat progenies (in females that were exposed to alcohol before mating with intact males), are considerably correctable with the use of the zinc sulfate water solution (when administered in to the rat females during pregnancy and lactation).
4. The alcohol exposure (in rat females - before mating with intact males) significantly affects the absolute concentrations of macro- and microelements within the organs and brain tissue samples of the rat mature progenies.
5. The chemical elements proportions within the organs and brain structures of the mature rat progenies are less prone to deviation, then the absolute values (chemical elements concentrations), including the groups of rats that were given zinc sulfate as a correction treatment.
Practical significance
The results of the performed investigation can be used for educational purposes in medical and pharmaceutical higher educational institutions, as well as in the biology and chemistry teacher training departments (for providing information about the pre-natal alcohol consumption’s great influence on the health conditions of children). The detected cases of elemental disproportions within the organs and brain structures of the rat mature progenies (alcoholisation before mating with intact males) show signs of the development of the metabolism impairments and physiological disturbances (that take place in the postnatal period), which serve as an evidence of the strict necessity for seeking ways to correct (ante- or postnatally) the negative consequences of the alcohol intoxication in female rats before they got pregnant. The perspectives of such a therapeutic approach are still to be researched.
Zinc sulfate and other zinc formulations may be considered as a treatment medication for correcting the negative consequences (found in internal organs and brain structures of the mature progenies) of the short-time periods of alcohol intoxication in rat females. The administration of the zinc-containing formulations during pregnancy and breast-feeding with episodes of alcohol exposure to the rat females before mating enhances the effectiveness of the compensatory and adaptive reactions that provide the intactness of the structural homeostasis in the central nervous system and the whole organism at a molecular level.
Thesis approbation
The materials of the research work were reported during the VIII ISTERH conference (The International Society for Trace Element Research in Humans) «Trace Elements in Diet, Nutrition and Health: Essentiality and Toxicity» (Crete, Greece, October, 2007); during the All-Russian Scientific Research Conference with international cooperative participation «Physiologic and Hygienic problems of the human ecology» (Belgorod, May 2007); during the 1st Russian national Narcology Congress (Moscow, November, 2009); the materials of the research work were reported to task group of the Federal State Institution “National Scientific Narcology Center”, (Moscow, April, 2010); the materials of the research work were reported during the 4th International Symposium on Trace Elements and Minerals in Medicine and Biology) (Saint Petersburg, June 2010).
The results of the research work –publications
The results of the research work were issued in 10 separate articles, among which 3 are published in journals licensed by the State Commission for Academic Degrees and Titles (Ministry of Education and Science of the Russian Federation). The results of the research work were mentioned in 2 chapters of the multi-author book.
Implementation of the research results
The results of the research work were implemented into the investigational activities of the Institute of Bioelementology, a subunit of the State Eductational Institution «Orenburg State University»; the implementation has also been performed in the Federal State Scientific Institution «The Institute of Toxicology», a subunit of the Federal Bio-Medical Agency of the Russian Federation; the results of the research work were implemented into the educational process (the Department of Nutriciology and Bioelementology of the State Educational Institution «Orenburg State University», the Department of Normal Physiology of the Peoples’ Friendship University of Russia. The results of the researched work provided a rationalization of adding the zinc asparaginate to the specially designed “Universitetskiy” drinking product (–manufactured by “Zhivaya voda” Ltd. (Orenburg).
The structure and the extent of the thesis work
The thesis work includes 140 pages of typed text, including the following chapters: introduction, background, materials and methods (that were used for the research work), results summary and discussions of the obtained data, conclusions, applicational recommendations and references list. The illustrative material includes 18 figures and 16 tables. The reference list contains 193 bibliographic sources (72- national and 121 foreign).
Research work: Contents
Investigative techniques
The research work has been performed on 227 random breed white rats: 32 females, 10 males, 185 – progenies.
The experimental animals were held in the bioclinics of the Institute of Bioelementology (a subunit of the State Eductational Institution «Orenburg State University» (director, M.D., professor А.V. Skalnyi) with the use of the usual light status (temperature +20-24°С). The basic food source for the experimental animals was – № 1cubed feed.
At the beginning of the experiment all the animals (32 rat females) were divided into 3 groups. The 1st group received 15% ethanol solution as the only source of liquid during 20 days (average values - every rat female was exposed to 13.3 g/kg of ethanol daily). On the day before mating with intact males the rat females of the 1st group were give usual water. The 2nd group of experimental animals received ethanol just like the 1st group, and on the day before mating with intact males the rat females were consuming a 0.003% water solution of zinc sulfate (average values: every animal received 0.05-0.07 µg/kg of Zn daily). The rat females of the 2nd group were receiving a water solution of Zn sulfate for the next 45 days after mating (i.e. for the whole bearing and breast-feeding period). The 3rd group of experimental animals received only water for the whole experimental period.